白血病患者p16基因失活机制研究.doc

白血病患者p16基因研究 【摘要】目的：了解白血病患者p16基因启动子甲基化，探讨在白血病发生发展中的作用及临床应用价值。方法：采用聚合酶链反应（polymerase Chain Reaction, PCR）、聚合酶链反应-单链构象多态性（polymerase chain reaction-single strand conformation polymorphism, PCR-SSCP）以及巢式甲基化特异性PCR法（nested methylation-specific PCR,nMSP），对57例白血病患者进行p16基因纯合性缺失、点突变及启动子甲基化研究。结果：在受检的57例白血病患者中，共有6例发生p16基因纯合性缺失10.5%）；5例受检者均未见点突变发生；受检的5例患者中p16基因甲基化，ALL组最高（41.2%），其次是AML（33.3%）、CLL（16.7%）和CML（1.3%）。血病发生过程中表观遗传学机制和遗传学机制相互作用，[关键词] 白血病p16基因甲基化 p16 gene inactivation in Leukemia XIAO Yun, LIU Yong-mei, JIANG Lin, RAN Lian-hui（Department of Clinical Laboratory,Guiyang Medical University,Guiyang 550004,China） [ABSTRACT] Objective: To investigate the deletion, the point mutation and the hypermethylation of promoter of p16 in leukemia and to discuss the mechanism of p16 gene inactivation in leukemia and its clinical sigificance. Methods: The deletion, point mutation and hypermethylation status of the promoter-associated CpG islands of p16 were analyzed by polymerase Chain Reaction(PCR), PCR-single strand conformation polymorphism(PCR-SSCP) and nested methylation-specific PCR(nMSP） in leukemia (AML) at diagnosis. Results: Among 57 patients, only 6 cases deletions（10.5%） were found. The mutation were not observed in any case. Methylation analysis of p16 gene promoter revealed hypermethylation of CpG islands in 16/51 cases (31.4%). Hypermethylation of p16 was observed in 41.2% of ALL, 33.3% of AML, 16.7% CLL and 14.3% of CML. Conclusion: Both epigenetic and genetic alternations were involved in leukemia formation. The genetic alterations including the deletion and the mutation of p16 gene were rare events whereas hypermethylation of p16 was the common event in leukemia, mainly in the acute leukemia. [Key words] Leukemia; p16; Deletion; Point mutation; Methylation 抑癌基因失活与肿瘤发生密切相关。经典遗传学机制认为核酸是遗传的分子基础，与DNA序列改变有关的纯合性缺失和基因突变是基因失活的主要形式。近年[1]。 p16基因重要的抑癌基因，通过抑制CDK4/6（Cyclin Dependent Kinases,细胞周期蛋白依赖性激酶）活性，使细胞周期停滞于G1/S期，对细胞周期起着负性调控作用。p16基因失活见于多种人类肿瘤，造血系统恶性肿瘤有关p16基因失活方式及发生率的报道结果较